ANESTHETIC EFFECTS ON SARCOPLASMIC RETICULUM CA++ ATPASE

描述：(改编自申请人的摘要)初稿 已找到申请人的全身麻醉药和局部麻醉药的实验 对心肌细胞Ca-ATPase活性及寡聚态的影响 骨骼肌和心肌的肌浆网(SR)。这些 结果与四聚体及更大分子的解释一致 低聚物比Ca-ATPase的单体和二聚体活性低得多。这个 长期目标是阐明这些疾病的分子机制。 麻醉药对Ca-ATPase的影响及其相关性 对全身麻醉药的心肌抑制作用和 局麻药的抗心律失常作用。CA-ATPase将被标记为 用共价磷光探针和时间分辨磷光 光谱学将被用来估计Ca-ATPase的寡聚状态 在心肌和骨骼肌的SR膜上。《将军》的影响 局麻药对大鼠心肌细胞低聚体状态及Ca-ATPase活性的影响 心脏和骨骼的SR将被测定。用AT-1的SR进行实验 细胞，一种缺乏磷蛋白的心房肿瘤细胞系，将有助于 确定磷蛋白是否对截然不同的 心脏和骨骼肌质网Ca-ATPase对全麻药的反应。这个 申请者将确定麻醉剂对患者反应的影响 心肌细胞Ca-ATPase对其两种主要成分的寡聚状态和活性 磷蛋白的生理调节、钙和磷酸化以及副产物 反过来说。在骨骼和心脏SR中，他们都会询问是否 局麻药促进血管内皮细胞大分子寡聚体形成的作用 CA-ATPase依赖于带正电的季铵盐基 大多数局部麻醉药。麻醉药对麻醉反应的影响 心肌和骨骼的Ca-ATPase对pH的改变，反之亦然 调查过了。文献资料博大精深，知之甚少。 PH值对吸入性心肌肌浆网Ca-ATPase反应的影响 被提议的实验旨在解释的麻醉剂。原子 力显微镜将提供一种独立的方法来证实或驳斥 麻醉诱导的Ca-ATPase寡聚体状态的变化 由先前的时间分辨磷光的结果表明 光谱学。

DESCRIPTION: (Adapted from the applicant's abstract) In preliminary experiments of the applicant, general and local anesthetics have been found to influence the activity and the oligomeric state of the Ca-ATPase of sarcoplasmic reticulum (SR) of both skeletal and cardiac muscle. These results are consistent with the interpretation that tetrameres and larger oligomers are much less active than monomers and dimers of Ca-ATPase. The long term objective is to elucidate the molecular mechanisms of these effects of anesthetics on the Ca-ATPase and to understand their relevance to the myocardial depressive effects of general anesthetics and the antiarrhythmic effects of local anesthetics. Ca-ATPase will be labeled with a covalent phosphorescent probe, and time resolved phosphorence spectroscopy will be used to estimate the oligomeric state of the Ca-ATPase in SR membranes of cardiac and skeletal muscle. The influences of general and local anesthetics on the oligomeric state and activity of Ca-ATPase of cardiac and skeletal SR will be determined. Experiments with SR from AT-1 cells, an atrial tumor cell line deficient in phospholamban, will help to determine whether phospholamban is responsible for the very different responses of cardiac and skeletal SR Ca-ATPase to general anesthetics. The applicants will determine the influence of anesthetics on the responses of the oligomeric state and activity of cardiac Ca-ATPase to its two major physiologic regulators, Ca and phosphorylation of phospholamban, and vice versa. In both skeletal and cardiac SR, they will inquire whether the effect of local anesthetics to promote the formation of large oligomers of Ca-ATPase depends upon the positively charged quaternary ammonium moiety of most local anesthetics. The influence of anesthetics on the response of cardiac and skeletal Ca-ATPase to altered pH, and vice versa, will be investigated. The literature documents profound and poorly understood influences of pH on the responses of cardiac SR Ca-ATPase to inhalational anesthetics that the proposed experiments are designed to explain. Atomic force microscopy will provide an independent method to confirm or refute the anesthetic induced changes in oligomeric state of Ca-ATPase that are indicated by the previous results with time resolved phosphorescence spectroscopy.